Compounds and methods for treating nematode infections

ABSTRACT

The present application relates to the treatment of nematode infections. For example, the application relates to the use of compounds of Formula (I) and/or (II) as defined herein for treatment of a nematode infection or a disease, disorder or condition arising from a nematode infection:

RELATED APPLICATIONS

The present application is a continuation-in-part of co-pending U.S. patent application Ser. No. 16/565,917 filed on Sep. 10, 2019, which claims the benefit of priority of U.S. provisional patent application No. 62/730,286 filed on Sep. 12, 2018, the contents of each of which are incorporated herein by reference in their entirety.

FIELD

The present application relates to the treatment of nematode infections. For example, the application relates to the use of compounds of Formula (I), and/or Formula (II) as defined herein for treatment of a nematode infection or a disease, disorder or condition arising from a nematode infection.

INTRODUCTION

The burden of parasitic nematodes on humanity is severe. The WHO estimates that over two billion people are infected with at least one parasitic nematode species. Chronic infection can cause dietary deficiency, anemia, developmental delay, elephantiasis, blindness, and death. Human infection in the west is increasing, coincident with a warming climate and the movement of sub-tropical species northward. Intestinal nematode infections alone are responsible for an estimated disease burden of 3.4 million disability-adjusted life-years. Furthermore, nematode infestation of food increases costs and contributes to malnutrition. Nematodes have evolved widespread resistance to nearly every anthelmintic (anti-worm) drug on the market. Hence, there is a dire need for the development of new compounds that kill parasitic worms.

Most anthelmintics disrupt the worm's nervous system, which ultimately allows the mammalian host to clear the infection. For example, key targets of ivermectin are glutamate-gated chloride channels of the nervous system. Ivermectin agonizes these channels and hyperpolarizes the cell, in turn leading to paralysis. Similarly, levamisole agonizes nicotinic acetylcholine receptors leading to depolarization of muscles and, in turn, paralysis.

SUMMARY

It has been shown herein that compounds of Formula (I) and/or (II) provide inhibitory activity against species of nematodes such as Cooperia oncophora, Haemonchus contortus, and Caenorhabditis elegans.

Accordingly, in one embodiment, the present application includes methods and uses for treating or preventing a nematode infection comprising administering an effective amount of a compound of Formula (I)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof wherein L is C₀₋₄ alkylene;

R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and

R² is H, NO₂, Cl, F or CN.

In one embodiment, the present application includes methods and uses for treating or preventing a nematode infection comprising administering an effective amount of a compound of Formula (II)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof.

In some embodiments, the present application includes methods and uses of treating or preventing a disease, disorder or condition arising from a nematode infection comprising administering an effective amount a compound of Formula (I)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof wherein L is C₀₋₄ alkylene; R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and R² is H, NO₂, Cl, F or CN.

In some embodiments, the present application includes methods and uses of treating or preventing a disease, disorder or condition arising from a nematode infection comprising administering an effective amount of a compound of Formula (II)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof.

In some embodiments, the present application includes pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a compound of Formula (I)

and/or a pharmaceutically acceptable salt and/or solvate thereof wherein L is C₀₋₄ alkylene; R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and R² is H, NO₂, Cl, F or CN; and wherein the compound of Formula (I) is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the present application includes pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a compound of Formula (II)

and/or a pharmaceutically acceptable salt and/or solvate thereof, wherein the compound of Formula (II) is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the present application includes uses of pharmaceutical compositions comprising a compound of Formula (I) and/or (II) for treating or preventing a nematode infection or a disease, a disorder, or a condition arising from a nematode infection in a subject in need thereof.

In some embodiments, the present application includes nematicidal compositions comprising a carrier and a compound of Formula (I)

and/or a salt and/or solvate thereof wherein L is C₀₋₄ alkylene; R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and R² is H, NO₂, Cl, F or CN; and wherein the compound of Formula (I) is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the present application includes nematicidal compositions comprising a carrier and a compound of Formula (II)

and/or a salt, and/or solvate thereof wherein the compound of Formula (II) is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the present application includes uses of the nematicidal composition described herein for treating or preventing a nematode infection or a disease, a disorder, or a condition arising from a nematode infection in a subject in need thereof.

In some embodiments, the present application includes methods of treating or preventing a nematode infection comprising administering an effective amount of a nematicidal composition described herein to a subject in need thereof.

Other features and advantages of the present application will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating embodiments of the application, are given by way of illustration only and the scope of the claims should not be limited by these embodiments, but should be given the broadest interpretation consistent with the description as a whole.

DRAWINGS

The embodiments of the application will now be described in greater detail with reference to the attached drawings in which:

FIG. 1 shows movement defects induced by exemplary compounds including compound Ia. Panel A shows a negative control healthy C. elegans worm without drug. Panel B shows the “coiler defect” phenotype of a C. elegans worm in presence of exemplary Compound Ia.

FIG. 2 shows impact of exemplary compound Ia on HEK293 cell proliferation compared to a positive control where dashed line shows data for exemplary compound Ia.

FIG. 3 shows drug-induced paralysis suppression in C. elegans in the presence of exemplary compound Ia. Circle (●) indicates negative control where no drug is used. Square (▪) shows positive control with Aldicarb, a paralysis-inducing drug. Downward-facing triangle (▾) indicates exemplary compound Ia used in combination with Aldicarb. Upward-facing triangle (▴) indicates exemplary compound Ia used alone.

DESCRIPTION OF VARIOUS EMBODIMENTS I. Definitions

Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present application herein described for which they are suitable as would be understood by a person skilled in the art.

In understanding the scope of the present application, the term “comprising” and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and/or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and/or steps. The foregoing also applies to words having similar meanings such as the terms, “including”, “having” and their derivatives.

The term “consisting” and its derivatives, as used herein, are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and/or steps, but exclude the presence of other unstated features, elements, components, groups, integers and/or steps.

The term “consisting essentially of”, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and/or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and/or steps.

Terms of degree such as “substantially”, “about” and “approximately” as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies.

As used in this application, the singular forms “a”, “an” and “the” include plural references unless the content clearly dictates otherwise. For example, an embodiment including “a compound” should be understood to present certain aspects with compound or two or more additional compounds.

In embodiments comprising an “additional” or “second” component, such as an additional or second compound, the second component as used herein is chemically different from the other components or first component. A “third” component is different from the other, first, and second components, and further enumerated or “additional” components are similarly different.

The term “and/or” as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that “at least one of” or “one or more” of the listed items is used or present. The term “and/or” with respect to pharmaceutically acceptable, salts and/or solvates thereof means that referenced compounds exist as individual salts or hydrates, as well as a combination of, for example, a salt of a solvate of a compound.

The term “suitable” as used herein means that the selection of the particular compound or conditions would depend on the specific synthetic manipulation to be performed, the identity of the molecule(s) to be transformed and/or the specific use for the compound, but the selection would be well within the skill of a person trained in the art. All process/method steps described herein are to be conducted under conditions sufficient to provide the product shown. A person skilled in the art would understand that all reaction conditions, including, for example, reaction solvent, reaction time, reaction temperature, reaction pressure, reactant ratio and whether or not the reaction should be performed under an anhydrous or inert atmosphere, can be varied to optimize the yield of the desired product and it is within their skill to do so.

The present description refers to a number of chemical terms and abbreviations used by those skilled in the art. Nevertheless, definitions of selected terms are provided for clarity and consistency.

The term “alkyl” as used herein, whether it is used alone or as part of another group, means straight or branched chain, saturated alkyl groups. The number of carbon atoms that are possible in the referenced alkyl group are indicated by the prefix “C_(n1-n2)”. For example, the term C₁₋₁₀alkyl means an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.

The term “alkylene”, whether it is used alone or as part of another group, means straight or branched chain, saturated alkylene group, that is, a saturated carbon chain that contains substituents on two of its ends. The number of carbon atoms that are possible in the referenced alkylene group are indicated by the prefix “C_(n1-n2)”. For example, the term C₁₋₆alkylene means an alkylene group having 1, 2, 3, 4, 5 or 6 carbon atoms.

The term “cycloalkyl,” as used herein, whether it is used alone or as part of another group, means a saturated carbocyclic group containing one or more rings. The number of carbon atoms that are possible in the referenced cycloalkyl group are indicated by the numerical prefix “C_(n1-n2)”. For example, the term C₃₋₈cycloalkyl means a cycloalkyl group having 3, 4, 5, 6, 7, or 8 carbon atoms.

The term “cycloalkenyl” as used herein, whether it is used alone or as part of another group, means cyclic, unsaturated alkyl groups. For example, the term C₃₋₈cycloalkenyl means a cycloalkenyl group having 3, 4, 5, 6, 7, or 8 carbon atoms and at least one double bond.

All cyclic groups, including phenyl, cycloalkyl and cycloalkenyl groups, contain one or more than one ring (i.e. are polycyclic). When a cyclic group contains more than one ring, the rings may be fused, bridged, spirofused or linked by a bond.

The term “benzofused” as used herein refers to a polycyclic group in which a benzene ring is fused with another ring.

A first ring being “fused” with a second ring means the first ring and the second ring share two adjacent atoms there between.

A first ring being “bridged” with a second ring means the first ring and the second ring share two non-adjacent atoms there between.

A first ring being “spirofused” with a second ring means the first ring and the second ring share one atom there between.

The term “substituted” as used herein means that the referenced atom contains at least one substituent group other that a hydrogen atom.

The term “nematode infection” as used herein refers to an invasion of cells or bodily tissues by a foreign undesirable nematode.

The term “anthelmintic” or “anthelmintics” as used herein refers to a group of antiparasitic drug used in the treatment and prevention of nematode infections.

As used herein, a compound with “anthelmintic activity” is a compound, which when tested, has measurable nematode-killing activity or results in sterility or reduced fertility in the nematodes such that fewer viable or no offspring result, or compromises the ability of the nematode to infect or reproduce in its host, or interferes with the growth or development of a nematode. The compound may also display nematode repellant properties. The term “pharmaceutically acceptable salt” means an acid addition salt or a basic addition salt suitable for, or compatible with, the treatment of subjects.

The term “pharmaceutically acceptable salts” embraces salts commonly used to form addition salts of free acids or free bases. The nature of the salt is not critical, provided that it is pharmaceutically-acceptable. Suitable pharmaceutically acceptable acid addition salts are prepared from an inorganic acid or an organic acid. Examples of such inorganic acids include, without limitation, hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuric and phosphoric acid. Examples of appropriate organic acids include, for example, aliphatic, cycloaliphatic, aromatic, arylaliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include, without limitation, formic, acetic, propionic, succinic, glycolic, gluconic, maleic, embonic (pamoic), methanesulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, pantothenic, benzenesulfonic, toluenesulfonic, sulfanilic, mesylic, cyclohexylaminosulfonic, stearic, algenic, β-hydroxybutyric, malonic, galactic, and galacturonic acid. Suitable pharmaceutically-acceptable base addition salts include, but are not limited to, metallic salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc or organic salts made from N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, lysine and procaine.

The term “solvates” as used herein refers to complexes formed between a compound and a solvent from which the compound is precipitated or in which the compound is made. Accordingly, the term “solvate” as used herein means a compound, or a salt a compound, wherein molecules of a suitable solvent are incorporated in the crystal lattice. Examples of suitable solvents are ethanol, water and the like. When water is the solvent, the molecule is referred to as a “hydrate”.

The term “pharmaceutically acceptable solvate” means a solvate suitable for, or compatible with, the treatment of subjects. For pharmaceutically acceptable solvates, a suitable solvent is physiologically tolerable at the dosage used or administered.

The term “compound(s) of the application” as used herein refers to a compound of Formula (I) or Formula (II) and/or a pharmaceutically acceptable salt, and/or solvate thereof.

The expression “disease, disorder or condition arising from a nematode infection” as used herein refers to any disease, disorder or condition that is directly or indirectly caused by the presence of a nematode infection in a subject.

The term “subject” as used herein includes plants, seeds, soil, and all members of the animal kingdom including mammals and birds, and their food. Thus, the methods of the present application are applicable to plant treatment, human therapy and veterinary applications.

When used, for example, with respect to the methods of treatment, uses, compositions and kits of the application, a subject, for example a subject “in need thereof” is a subject who has been diagnosed with, is suspected of having, may come in to contact with, and/or was previously treated for a nematode infection or a disease, disorder or condition arising from a nematode infection.

When used, for example, in respect to plant treatments, the compounds and/or compositions may be delivered by several means including pre-planting, post-planting and as a feed additive, drench, external application, pill or by injection.

The term “pharmaceutical composition” as used herein refers to a composition of matter for pharmaceutical use.

The term “pharmaceutically acceptable” means compatible with the treatment of subjects.

The term “pharmaceutically acceptable carrier” means a non-toxic solvent, dispersant, excipient, adjuvant or other material which is mixed with the active ingredient in order to permit the formation of a pharmaceutical composition, i.e., a dosage form capable of administration to a subject.

The term “nematicidal composition” as used here in refers to a composition of matter for managing one or more nematode infections.

The term “administered” as used herein means administration of an effective amount of a compound, including compounds of Formula (I) or (II), or a salt and/or solvate thereof, to a cell either in cell culture or in a subject.

As used herein, the term “effective amount” or “therapeutically effective amount” means an amount effective, at dosages and for periods of time necessary to achieve a desired result. For example, in the context of treating a nematode infection, or a disease, disorder or condition arising from a nematode infection, an effective amount of a compound is an amount that, for example, reduces the nematode infection compared to the nematode infection without administration of the compound. By “reducing the infection”, it is meant, for example, reducing the amount of the infectious agent in the subject and/or reducing the symptoms of the infection. The amount of a given compound or composition that will correspond to such an amount will vary depending upon various factors, such as the given compound or composition, the formulation, the route of administration, the type of condition, disease or disorder, the identity of the subject being treated, and the like, but can nevertheless be routinely determined by one skilled in the art.

The terms “to treat”, “treating” and “treatment” as used herein and as is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results include, but are not limited to, diminishment of extent of nematode infection, stabilization (i.e. not worsening) of the state of the nematode infection, preventing spread of the nematode infection, delay or slowing of infection progression, amelioration or palliation of the nematode infectious state, diminishment of the reoccurrence of nematode infection, diminishment, stabilization, alleviation or amelioration of one or more diseases, disorders or conditions arising from the nematode infection, diminishment of the reoccurrence of one or more diseases, disorders or conditions arising from the nematode infection, and remission of the nematode infection and/or one or more symptoms or conditions arising from the nematode infection, whether partial or total, whether detectable or undetectable. “To treat”, “treating” and “treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. “To treat”, “treating” and “treatment” as used herein also include prophylactic treatment. For example, a subject with an early nematode infection is treated to prevent progression, or alternatively a subject in remission is treated to prevent recurrence.

“Palliating” an infection, disease, disorder and/or condition means that the extent and/or undesirable manifestations of an infection, disease, disorder and/or condition are lessened and/or time course of the progression is slowed or lengthened, as compared to not treating the infection, disease, disorder and/or condition.

The term “prevention” or “prophylaxis” and the like as used herein refers to a reduction in the risk or probability of a subject becoming afflicted with a nematode infection and/or a disease, disorder and/or condition arising from a nematode infection or manifesting a symptom associated with a nematode infection and/or a disease, disorder and/or condition arising from a nematode infection.

II. Methods and Uses of the Application

Families of small molecule compounds have been identified that incapacitate parasitic nematodes, and advantageously show no genetic resistance. This suggest that resistance to these compounds will be less likely to develop in the wild.

The present application includes methods and uses for treating or preventing a nematode infection comprising administering an effective amount of a compound of Formula (I)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof wherein L is C₀₋₄ alkylene; R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and R² is H, NO₂, Cl, F or CN.

In one embodiment, the present application includes methods and uses for treating or preventing a nematode infection comprising administering an effective amount of a compound of Formula (II)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof.

In some embodiments, the present application includes methods and uses of treating or preventing a disease, disorder or condition arising from a nematode infection comprising administering an effective amount a compound of Formula (I)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof wherein L is C₀₋₄ alkylene; R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and R² is H, NO₂, Cl, F or CN.

In some embodiments, the present application includes methods and uses of treating or preventing a disease, disorder or condition arising from a nematode infection comprising administering an effective amount of a compound of Formula (II)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof.

In some embodiments, the present application includes uses of pharmaceutical compositions comprising a compound of Formula (I) and/or (II) for treating or preventing a nematode infection or a disease, a disorder, or a condition arising from a nematode infection in a subject in need thereof.

In some embodiments, the present application includes methods of treating or preventing a nematode infection comprising administering an effective amount of a nematicidal composition comprising a compound of Formula (I) and/or (II) described herein to a subject in need thereof.

In some embodiments, the present application includes uses of a nematicidal composition comprising a compound of Formula (I) and/or (II) described herein for treating or preventing a nematode infection or a disease, a disorder, or a condition arising from a nematode infection in a subject in need thereof.

In some embodiments, the L of compound of Formula (I) is CH₂.

In some embodiments, the L of compound of Formula (I) is absent, such that R¹ is directly bonded to the N atom.

In some embodiments, R¹ is C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with a C₁₋₄alkyl group. In some embodiments, R¹ is C₃₋₈cycloalkyl which is unsubstituted. In some embodiments, R¹ is C₃₋₈cycloalkyl which is substituted with a C₁₋₄alkyl group. In some embodiments, R¹ is C₃₋₈cycloalkenyl which is unsubstituted.

In some embodiments R¹ is unsubstituted C₅₋₇cycloalkyl.

In some embodiments, R² is H, CN or NO₂. In some embodiments, R¹ is CN or NO₂. In some embodiments, R² is CN. In some embodiments, R² is NO₂. In some embodiments, the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the compound of Formula (I) is

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the nematode infection is an infection of a nematode of a species selected from Cooperia oncophora, Haemonchus contortus, and Caenorhabditis elegans.

Treatment methods comprise administering to a subject one or more compounds of the application, and optionally consists of a single administration, or alternatively comprises a series of administrations. The length of the treatment period depends on a variety of factors, such as the severity of the infection, disease, disorder or condition, the age of the subject, the dosage of the one or more compounds of the application, the activity of one or more compounds of the application, or a combination thereof.

In an embodiment, the one or more compounds of the application are administered or used as soon as possible after exposure to the nematode. In an embodiment, the one or more compounds of the application are administered or used until treatment of the nematode infection, disease disorder or condition is achieved. For example, until complete elimination of the nematode is achieved, or until the number of nematode has been reduced to the point where the subject's defenses are no longer overwhelmed and can kill any remaining nematode.

In an embodiment, the methods of the present application comprise administering an effective amount of a compound or a composition of the application to a subject selected from humans, mammals, birds, vertebrates, plants, seeds, and soil.

In an embodiment, the uses of the present application of a compound or a composition of the application are in a subject selected from humans, mammals, birds, vertebrates, plants, seeds, and soil.

In some embodiments, the nematode infects plants and the nematicidal composition is administered to the soil or to plants. In some embodiments, the nematicidal composition is administered to soil before planting. In some embodiments, the nematicidal composition is administered to soil after planting. In some embodiments, the nematicidal composition is administered to soil using a drip system. In some embodiments, the nematicidal composition is administered to soil using a drench system. In some embodiments, the nematicidal composition is administered to plant roots or plant foliage (e.g., leaves, stems). In some embodiments the nematicide composition is tilled into the soil or administered in furrow. In some embodiments, the nematicidal composition is administered to seeds.

In some embodiments, the nematode parasite infects a vertebrate. In some embodiments, the nematicidal composition is administered to non-human vertebrate. In some embodiments, the nematicidal composition is administered to a human. In some embodiments, the nematicidal composition is formulated as a drench to be administered to a non-human animal. In some embodiments, the nematicidal composition is formulated as an orally administered drug. In some embodiments, the nematicidal composition is formulated as an injectable drug. In some embodiments, the nematicidal composition is formulated for topical applications such as pour-ons, or for the use in tags or collars.

In some embodiments, the methods of the application comprise administering a compound or a composition of the application through one or more means selected from pre-planting, post-planting, as a feed additive, a drench, an external application, a pill and by injection.

In some embodiments, the present application includes methods of reducing the viability or fecundity or slowing the growth or development or inhibiting the infectivity of a nematode using a compound or a composition of the application as described herein.

In some embodiments, the present application includes methods of reducing the viability or fecundity or slowing the growth or development or inhibiting the infectivity of a nematode using a compound or a composition of the application as described herein, the methods comprising administering a compound or a composition of the application to subject selected from a human, a mammal, a bird, a vertebrate in general, a plant, a seed, or soil. In some examples, the bird can be a domesticated fowl; the mammal can be a domesticated animal and/or livestock.

The dosage of the one or more compounds of the application, varies depending on many factors such as the pharmacodynamic properties thereof, the mode of administration, the age, health and weight/mass of the subject, the nature and extent of the symptoms, the frequency of the treatment and the type of concurrent treatment, if any, and the clearance rate in the subject to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. The one or more compounds of the application may be administered initially in a suitable dosage that may be adjusted as required, depending on the response.

Compounds can be tested for anthelmintic activity using methods known in the art. For example, the compound is combined with nematodes, e.g., in a well of microtiter dish, in liquid or solid media or in the soil containing the agent. Staged nematodes are placed on the media. The time of survival, viability of offspring, and/or the movement of the nematodes are measured. An agent with “anthelmintic or anthelminthic or antihelmthic activity” can, for example, reduce the survival time of adult nematodes relative to unexposed similarly staged adults, e.g., by about 20%, 40%, 60%, 80%, or more. In the alternative, an agent with “anthelmintic or anthelminthic or antihelminthic activity” may also cause the nematodes to cease replicating, regenerating, and/or producing viable progeny, e.g., by about 20%, 40%, 60%, 80%, or more. The effect may be apparent immediately or in successive generations.

III. Compositions of the Application

In some embodiments, the present application includes pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a compound of Formula (I)

and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof wherein: L is C₀₋₄ alkylene; R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and R² is H, NO₂, Cl, F or CN; and wherein the compound of Formula (I) is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the present application includes pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a compound of Formula (I)I

and/or a pharmaceutically acceptable salt and/or solvate thereof, wherein the compound of Formula (I)I is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the present application includes uses of pharmaceutical compositions comprising a compound of Formula (I) or (II) for treating or preventing a nematode infection or a disease, a disorder, or a condition arising from a nematode infection in a subject in need thereof.

In some embodiments, the present application includes a nematicidal composition comprising a carrier and a compound of Formula (I)

and/or a salt and/or solvate thereof wherein: L is C₀₋₄ alkylene; R¹ is phenyl, C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with one or two substituents independently selected from C₁₋₄alkyl, C₁₋₄fluoroalkyl and C₁₋₄alkyl substituted with Cl or CN; and R² is H, NO₂, Cl, F or CN; and wherein the compound of Formula (I) is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the present application includes a nematicidal composition comprising a carrier and a compound of Formula (II)

and/or a salt and/or solvate thereof wherein the compound of Formula (II) is present in an amount effective to treat a nematode infection in a subject in need thereof.

In some embodiments, the linker L of compound of Formula (I) is CH₂.

In some embodiments, the linker L of compound of Formula (I) is optional and absent, such that R¹ is directly bonded to the N atom.

In some embodiments, R¹ is C₃₋₈cycloalkyl or C₃₋₈cycloalkenyl, each of which is unsubstituted or substituted with a C₁₋₄alkyl group. In some embodiments, R¹ is C₃₋₈cycloalkyl which is unsubstituted. In some embodiments, R¹ is C₃₋₈cycloalkyl which is substituted with a C₁₋₄alkyl group. In some embodiments, R¹ is C₃₋₈cycloalkenyl which is unsubstituted.

In some embodiments R¹ is unsubstituted C₅₋₇cycloalkyl.

In some embodiments, R² is H, CN or NO₂. In some embodiments, R¹ is CN or NO₂. In some embodiments, R² is CN. In some embodiments, R² is NO₂.

In some embodiments, the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the compound of Formula (I) is

or a pharmaceutically acceptable salt and/or solvate thereof.

In some embodiments, the nematode infection is an infection of a nematode of the following non-limiting, exemplary genera: Caenorhabditis, Nippostrongyles, Anguina, Ditylenchus, Tylenchorhynchus, Pratylenchus, Radopholus, Hirschmanniella, Nacobbus, Hoplolaimus, Scutellonema, Rotylenchus, Helicotylenchus, Rotylenchulus, Belonolaimus, Heterodera, other cyst nematodes, Meloidogyne, Criconemoides, Hemicycliophora, Paratylenchus, Tylenchulus, Aphelenchoides, Bursaphelenchus, Rhadinaphelenchus, Longidorus, Xiphinema, Trichodorus, Paratrichodorus, Dirofiliaria, Onchocerca, Brugia, Acanthocheilonema, Aelurostrongylus, Anchlostoma, Angiostrongylus, Ascaris, Bunostomum, Capillaria, Chabertia, Cooperia, Crenosoma, Dictyocaulus, Dioctophyme, Dipetalonema, Dracunculus, Enterobius, Filaroides, Haemonchus, Lagochilascaris, Loa, Manseonella, Muellerius, Necator, Nematodirus, Oesophagostomum, Ostertagia, Parafilaria, Parascaris, Physaloptera, Protostrongylus, Setaria, Spirocerca, Stephanogilaria, Strongy hides, Strongylus, Thelazia, Toxascaris, Toxocara, Trichinella, Trichostrongylus, Trichuris, Uncinaria or Wuchereria. In some embodiments, the nematodes are of the genera Cooperia, Haemonchus, Caenorhabditis, Nippostrongyles, Dirofilaria, Onchocerca, Brugia, Acanthocheilonema, Dipetalonema, Loa, Mansonella, Parafilaria, Setaria, Stephanofilaria, Wucheria, Pratylenchus, Heterodera, Meloidogyne or Paratylenchus. In some embodiments the nemotodes are of the species Cooperia oncophora, Haemonchus contortus, Caenorhabditis elegans, Nippostrongyles brasiliensis, Ancylostoma caninum, Haemonchus contortus, Trichinella spiralis, Trichurs muris, Dirofilaria immitis, Dirofilaria tenuis, Dirofilaria repens, Dirofilari ursi, Ascaris suum, Toxocara canis, Toxocara cati, Strongyloides ratti, Parastrongyloides trichosuri, Heterodera glycines, Globodera pallida, Meloidogyne javanica, Meloidogyne incognita, Meloidogyne arenaria, Radopholus similis, Longidorus elongatus, Meloidogyne hapla or Pratylenchus penetrans.

In some embodiments, the nematode infection is an infection of a nematode of a species selected from Cooperia oncophora, Haemonchus contortus, Dirofilaria, Nippostrongyles brasiliensis and Caenorhabditis elegans.

In some embodiments, the subject is selected from humans, mammals, birds, vertebrates, plants, seeds, and soil.

In some embodiments, the nematicidal compositions further comprise one or more agricultural excipients.

In some embodiments, the nematicidal compositions further comprise one or more agriculturally acceptable excipients.

For example, in some embodiments, the nematicidal compositions further comprises an aqueous surfactant. Examples of surfactants that can be used include, Span 20, Span 40, Span 80, Span 85, Tween 20, Tween 40, Tween 80, Tween 85, Triton X 100, Makon 10, Igepal CO 630, Brij 35, Brij 97, Tergitol TMN 6, Dowfax 3B2, Physan and Toximul TA 15, and mixtures thereof.

In some embodiments, the nematicidal composition further comprises a permeation enhancer (e.g., cyclodextrin). In some embodiments, the nematicidal composition further comprises a co-solvent. Examples of co-solvents that can be used include ethyl lactate, methyl soyate/ethyl lactate co-solvent blends (e.g., Steposol), isopropanol, acetone, 1,2-propanediol, n-alkylpyrrolidones (e.g., the Agsolex series), a petroleum based-oil (e.g., aromatic 200) or a mineral oil (e.g., paraffin oil), or mixtures thereof. In some embodiments, the nematicidal composition further comprises another pesticide (e.g., nematicide, insecticide or fungicide) such as an avermectin (e.g., ivermectin), milbemycin, imidacloprid, aldicarb, oxamyl, fenamiphos, fosthiazate, metam sodium, etridiazole, penta-chloro-nitrobenzene (PCNB), flutolanil, metalaxyl, mefonoxam, and fosetyl-al, or mixtures thereof. Useful fungicides include, but are not limited to, silthiofam, fludioxonil, myclobutanil, azoxystrobin, chlorothalonil, propiconazole, tebuconazole and pyraclostrobin, or mixtures thereof. In some embodiments, the nematicidal composition may also comprise herbicides (e.g., trifloxysulfuron, glyphosate, halosulfuron) and other chemicals for disease control (e.g., chitosan).

In some embodiments, the application also includes a nematicidal feed for a non-human vertebrate including:

(a) a feed; and (b) a nematicidal composition, of the application.

In some embodiments, the feed is selected from: soy, wheat, corn, sorghum, millet, alfalfa, clover, and rye, and mixtures thereof. Also described are feeds that have been supplemented to include one or more of the compounds of the application. A nematicidal feed for a non-human vertebrate can comprise: (a) an animal feed; and (b) an effective amount of a nematicidal compound of the application.

In some embodiments, the pharmaceutical compositions further comprise one or more pharmaceutically acceptable excipients. Conventional procedures and ingredients for the selection and preparation of suitable pharmaceutical compositions are described, for example, in Remington's Pharmaceutical Sciences (2000-20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999.

For example, the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form is sterile and fluid to the extent that easy syringability exists.

In an embodiment, parenteral administration is by continuous infusion over a selected period of time. Solutions suitable for parenteral administration are prepared by known methods by a person skilled in the art. For example, the compounds of the application are prepared in water optionally mixed with a surfactant such as hydroxypropylcellulose. Dispersions are also prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

Compositions for nasal administration are conveniently formulated as aerosols, drops, gels or powders. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which take the form of a cartridge or refill for use with an atomising device. Alternatively, the sealed container is a unitary dispensing device such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve, which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it contains a propellant, which is, for example, a compressed gas such as compressed air or an organic propellant such as fluorochlorohydrocarbon. In an embodiment, the aerosol dosage forms take the form of a pump-atomizer.

Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, wherein the active ingredient is formulated with a carrier such as sugar, acacia, tragacanth, gelatin and/or glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

In another embodiment, compounds of the application are orally administered, for example, with an inert diluent or with an assimilable edible carrier, or they are enclosed in hard or soft shell gelatin capsules, or they are compressed into tablets, or they are incorporated directly with the food of a diet. For oral administration, the compounds of the application may be incorporated with excipients and used in the form of, for example, ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Oral dosage forms also include modified release, for example immediate release and timed-release, formulations. Examples of modified-release formulations include, for example, sustained-release (SR), extended-release (ER, XR, or XL), time-release or timed-release, controlled-release (CR), or continuous-release (CR or Contin), employed, for example, in the form of a coated tablet, an osmotic delivery device, a coated capsule, a microencapsulated microsphere, an agglomerated particle, e.g., molecular sieving type particles, or, a fine hollow permeable fiber bundle, or chopped hollow permeable fibers, agglomerated or held in a fibrous packet. In an embodiment, timed-release compositions are, formulated, as liposomes or those wherein the active compound is protected with differentially degradable coatings, such as by microencapsulation, multiple coatings, etc. Liposome delivery systems include, for example, small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. In an embodiment, liposomes are formed from a variety of lipids, such as cholesterol, stearylamine or phosphatidylcholines.

It is also possible to freeze-dry the compounds of the application and use the lyophilizate obtained, for example, for the preparation of products for injection.

The compounds of the application are either commercially available or may be prepared using methods known in the art.

The formation of a desired compound salt is achieved using standard techniques. For example, the neutral compound is treated with an acid or base in a suitable solvent and the formed salt is isolated by filtration, extraction or any other suitable method.

The formation of solvates will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. The selection of suitable conditions to form a particular solvate can be made by a person skilled in the art.

The following non-limiting examples are illustrative of the present application. As is apparent to those skilled in the art, many of the details of the examples may be changed while still practicing the methods, compositions and kits described herein.

IV. Methods of Preparation

The compounds of the application are either commercially available or may be prepared using methods known in the art. For example, in some embodiments, compounds of Formula (I) are prepared as shown in Scheme I:

Therefore compounds of Formula (I), wherein R² is as defined in Formula (I), are reacted with excess amounts of piperazine (B) in the presence of an inorganic base at elevated temperatures to provide compounds of Formula C. Compounds of Formula C are then reacted with compounds of Formula D, wherein LG is a leaving group and R¹ is as defined in Formula (I), in the presence of an organic base, at elevated temperatures, to provide compounds of Formula (I).

The formation of a desired compound salt is achieved using standard techniques. For example, the neutral compound is treated with an acid in a suitable solvent and the formed salt is isolated by filtration, extraction or any other suitable method.

The formation of solvates will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. The selection of suitable conditions to form a particular solvate can be made by a person skilled in the art.

The following non-limiting examples are illustrative of the present application. As is apparent to those skilled in the art, many of the details of the examples may be changed while still practicing the methods, compositions and kits described herein.

EXAMPLES Example 1: Preparation of Compounds

Materials and Methods

Unless otherwise stated, all reactions were carried out under an atmosphere of dry argon, using glassware that was either oven (120° C.) or flame-dried. Work-up and isolation of compounds was performed using standard benchtop techniques. All commercial reagents were purchased from chemical suppliers (Sigma-Aldrich, Combi-Blocks, or Alfa Aesar) and used without further purification. Dry solvents were obtained using standard procedures (dichloromethane and acetonitrile were distilled over calcium hydride). Reactions were monitored using thin-layer chromatography (TLC) on EMD Silica Gel 60 F254 plates. Visualization was performed under UV light (254 nm) or using potassium permanganate (KMnO₄) stain. Flash column chromatography was performed on Siliaflash P60 40-63 μm silica gel purchased from Silicycle.

NMR characterization data was obtained at 293K on a Varian Mercury 300 MHz, Varian Mercury 400 MHz, Bruker Advance III 400 MHz, Agilent DD2 500 MHz equipped with a 5 mm Xses cold probe or Agilent DD2 600 MHz. ¹H spectra were referenced to the residual solvent signal (CDCl₃=7.26 ppm, DMSO-d₆=2.50 ppm). ¹³C{¹H} spectra were referenced to the residual solvent signal (CDCl₃=77.16 ppm, DMSO-d₆=39.52 ppm). Data for ¹H NMR are reported as follows: chemical shift (δ ppm), multiplicity (s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet), coupling constant (Hz), integration. NMR spectra were recorded at the University of Toronto Department of Chemistry NMR facility (http://www.chem.utoronto.ca/facilities/nmr/nmr.html).

Infrared spectra were recorded on a Perkin-Elmer Spectrum 100 instrument equipped with a single-bounce diamond/ZnSe ATR accessory in the solid state and are reported in wavenumber (cm⁻¹) units.

High resolution mass spectra (HRMS) were recorded at the Advanced Instrumentation for Molecular Structure (AIMS) in the Department of Chemistry at the University of Toronto (https://www.chem.utoronto.ca/chemistry/AIMS.php).

High-performance liquid chromatography (HPLC) analyses were performed on an Agilent 1100 series instrument containing a Phenomenex XDB-C18 column (1.8 μm, 50×4.6 mm) and run at room temperature. A linear gradient starting from 5% acetonitrile and 95% water (0.1% formic acid) to 95% acetonitrile and 5% water (0.1% formic acid) over 4 minutes followed by elution for 5 minutes at 95% acetonitrile and 5% water (0.1% formic acid) was used. The flow rate was set to 1.0 mL/min, with UV detection at 254 and 280 nm.

General Procedure A

The procedure was modified from literature (Sokoloff, P.; Pilon, C.; Mann, A.; Schoenfelder, A.; Garrido, F. 3,4-dihydro-2-naphthamide derivatives as selective dopamine D3 ligands. European Patent Application 05290156.8, Jul. 26 2006). To a round-bottom flask at room temperature were added piperazine (1.38 g, 16 mmol, 4.0 equiv), potassium carbonate (1.11 g, 8 mmol, 2 equiv), dimethyl sulfoxide (4.7 mL) then the corresponding fluorobenzene derivative (4 mmol, 1 equiv) and the mixture was stirred at 100° C. Once the reaction was complete as indicated by TLC (approximately 22 h), the reaction mixture was diluted with ethyl acetate, transferred to a separatory funnel and washed with water (3 times) then saturated sodium chloride. The organic phase was dried with Na₂SO₄, filtered, then concentrated on a rotary evaporator and the substituted 1-phenylpiperazine (C) was obtained. The crude solid was used in the next step without further purification.

General Procedure B

To a round-bottom flask were added C (0.5 mmol, 1.0 equiv), dry acetonitrile (1 mL), triethylamine (0.14 mL, 1 mmol, 2 equiv) then the alkyl halide wherein X is a halide (D, 0.5 mmol, 1.0 equiv) and the mixture was stirred at reflux until completion as indicated by TLC (approximately 24 h). The mixture was diluted with ethyl acetate, transferred to a separatory funnel and washed with water (3 times) then saturated sodium chloride. The organic phases were combined and dried with Na₂SO₄, filtered, and concentrated on a rotary evaporator. The residue was purified by column chromatography with the indicated eluent and the alkylated arylpiperazines (I) were obtained.

Preparation of cyclopentylmethyl 4-methylbenzenesulfonate

The procedure was modified from literature (Shi, W.; Nacev, B. A.; Aftab, B. T.; Head, S.; Rudin, C. M.; Liu, J. O. J. Med. Chem. 2011, 54, 7363). To a round-bottom flask were added cyclopentylmethanol (0.087 mL, 0.8 mmol, 1.0 equiv) and dry dichloromethane (1.6 mL) and the flask was submerged in an ice-water bath. Triethylamine (0.17 mL, 1.2 mmol, 1.5 equiv) was added to the flask followed by 4-(dimethylamino)pyridine (48.9 mg, 0.4 mmol, 0.5 equiv) then p-toluenesulfonyl chloride (168 mg, 0.88 mmol, 1.1 equiv) and the mixture was warmed to room temperature and stirred until completion as indicated by TLC (approximately 3 h). Dichloromethane and water were added, then the mixture was transferred to a separatory funnel and the layers were separated. The organic phase was washed with water (2 times) followed by saturated sodium chloride, then dried over Na₂SO₄, filtered, and concentrated on a rotary evaporator. The residue was purified by column chromatography (19/1 v/v pentanes/ethyl acetate) and cyclopentylmethyl 4-methylbenzenesulfonate (D(a), 59% yield) was obtained as a colourless oil. The spectral data was in accordance with literature.

Example 2 Screening of Compounds

Because of their dependence on a host, parasitic nematodes are difficult to culture, maintain and sustain. Therefore, to screen for antithelmintic compounds, the non-parasitic nematode, C. elegans, was used as a model system, then compound leads were investigated in parasitic nematodes.

Molecules that can modulate the egg-laying rate of the C. elegans (which can be done in medium-throughput fashion) were identified first and then the hits' ability to disrupt worm locomotion was examined using lower-throughput techniques. Egg-laying is a convenient system with which to interrogate neuromuscular control because it's digital output (an egg is either laid or not) can be easily quantified. Wild type hermaphrodites carry 10-15 eggs in their uterus and lay ˜one egg per hour in the liquid (control) buffer used for liquid worm culture. Egg-laying is principally regulated by cholinergic and serotonergic signaling via conserved receptors.

To identify egg-laying (Egl) modulators, a medium-throughput line-scanning microscope capable of imaging a 96 well plate in just over 2 minutes was used. From the resulting images, image analysis software to count the number of eggs laid per adult per hour in each well was used. Small molecules at a final concentration of 60 μM were screened, which is a technically convenient concentration where most molecules stay in solution. This may be considered a high concentration for most screening platforms, but is suitable for C. elegans because of the nematode's robust xenobiotic defenses.

A collection of 486 synthetic small molecules that have previously shown some effect on worm growth was screened. These molecules are from Chembridge Inc., are drug-like in their physico-chemical properties, and have relatively uncharacterized bioactivity. To identify Egl-stimulators, small molecules were screened in a benign control buffer. To identify Egl-inhibitors, molecules were screened in the background of 2.5 mM serotonin plus 7.7 mM nicotine, which is a cocktail that was empirically determined to induce robust egg-laying in liquid media. Molecules that reproducibly (p<0.05) modulate egg-laying two fold greater than control conditions were considered ‘stimulators’, and molecules that modulate egg-laying two fold less than stimulating conditions were considered ‘inhibitors’.

In the above-described assay, 31 stimulators and 33 inhibitors of egg-laying were identified. A structural comparison of these hits using a Tanimoto™/FP2 similarity cutoff of 0.55 or greater revealed that many of the Egl-modulators belong to structurally-similar families. A network was generated using a Tanimoto™ chemical similarity cut-off of 55%. Molecules that share structural similarity typically modulate egg laying similarly, suggesting a shared mechanism of action.

The ability of the 64 Egl hits to modulate the normal sinusoidal locomotion of wild type C. elegans was then examined. The animal's behaviour over the course of several hours was manually monitored while the animal moved atop a solid agar substrate containing the small molecule. Many different genes govern the sinusoidal locomotion of wild type C. elegans, but their mutation can lead to qualitatively distinct locomotory phenotypes. Hence, a distinct locomotory response to a small molecule can be suggestive of its mechanism of action.

Compared to the normal phenotype of the worm as shown in FIG. 1 panel A, 8 molecules that induce simultaneous anterior and posterior contraction (called a ‘rubber-band’ phenotype) were identified; 3 molecules that cause animals to turn along their circumferential axis (ca a ‘shaker’ phenotype) were identified; 3 molecules that cause the animal to coil like a snake (called a ‘coiler’ phenotype) were identified; 2 molecules that induce jerky uncoordination (called jerky-unc) were identified; 3 molecules that disrupt reverse locomotion were identified; and 3 that induce paralysis were identified.

Compound Ia induced a strong coiler phenotype (FIG. 1 panel B). Previous phylogenetic bioactivity analysis suggests that the activity of compound Ia might be restricted to nematodes. The coiler phenotype that is induced by compound Ia is consistent with a decrease in acetylcholine (ACh) levels at the synaptic cleft.

Example 3 Cytotoxicity of Exemplary Compound Ia

The cytotoxicity of exemplary compound Ia was investigated by measuring HEK293 cell proliferation in a dose-response analysis and no effect was found on HEK293 cell proliferation by exemplary compound Ia as shown in FIG. 2 . A cytotoxic formulation was used as positive control. Experiments were performed in triplicates on different days. Cell viability was assess using a standard Celltiter Blue™ Cell Viability Assay (Promega). Compounds were added to cells in 96-well plates at time zero and cell viability assessed in 48 hours. Error bars indicate standard deviation.

Example 4 Compounds of Formula (I) Induce Coiler Phenotype

Three structurally-related molecules (compound Ia, compound Ie and compounds Ic) induce a ‘severe’ snake-like coiler phenotype and paralysis (FIG. 1 , panel B). Compound Id is a forth structurally-related molecule that induces a ‘jerky-unc’ phenotype. A detailed phenotypic analyses suggests that the animals exposed to these molecules phenotypically transit over time from jerky-unc to being coiled to being paralyzed, with exemplary compound Ia being the prototypical example.

Forward genetic screens were performed for mutants that can resist the early larval lethal effects of compound Ic and compound Id. Over 200,000 F1 animals and over 100,000 F2 animals from randomly mutagenized parents/grandparents were screened. No mutants that could resist the lethal effects of compound Ic and compound Id. This suggests that viable resistance may be difficult to achieve against this class of nematicide in the wild.

Example 5 Mechanism of Action: Compounds of Formula (I) Likely Inhibits Vesicular Acetylcholine Transporter (VAChT)

Two observations raised the possibility that compounds of Formula (I) reduce acetylcholine (ACh) signalling. First, many mutant genes that yield a strong coiler phenotype are associated with disrupted cholinergic motorneuron development and/or with disrupted ACh production, release or reception. Second, the compounds of Formula (I) have structural similarity to vesamicol, which is a research tool that inhibits the vertebrate vesicular acetylcholine transporter (VAChT) (Table 1). VAChT transports cytoplasmic ACh into synaptic vesicles in the presynaptic membrane of all animals. Vesamicol is not useful as a medicine because it disrupts ACh signalling and is therefore toxic to vertebrates. In C. elegans, VAChT is encoded by unc-17, whose reduction-of-function phenotype is ‘severe’ coiling. It was therefore hypothesized that exemplary compound Ia inhibits UNC-17.

Exemplary compound Ia's inhibitory activity on UNC-17 was tested in several ways. First, the phenotype of exemplary compound Ia was compared to that of vesamicol. Vesamicol induces jerky-unc and coiling, but only at concentrations close to its limit of solubility on solid substrate. This analysis reveals that exemplary compound Ia is more than 30 times more potent than vesamicol for inducing locomotory defects in worms.

Whether exemplary compound Ia can suppress the paralysis that is induced by aldicarb was tested. Aldicarb inhibits acetylcholinesterase, which normally catabolizes ACh at the synaptic cleft. Exemplary compound Ia's EC₅₀ for suppression of Aldicarb activity is 4.806 μM. Aldicarb treatment therefore results in excess ACh and paralyzes the animal due to excess muscle contraction. If exemplary compound Ia inhibits UNC-17 and results in lower levels of ACh at the synaptic cleft, then exemplary compound Ia should reduce aldicarb-induced paralysis. Indeed, an obvious reduction of aldicarb-induced paralysis by exemplary compound Ia was found (p<0.001) (FIG. 3 ).

Third, the interaction between exemplary compound Ia and various mutant strains of unc-17 was explored. If exemplary compound Ia inhibits UNC-17, then unc-17 reduction-of-function alleles should be hypersensitive to exemplary compound Ia. Indeed, the unc-17 hypomorphic alleles e245, e327, and e795 have heightened sensitivity to exemplary compound Ia (p<0.001). Previous work demonstrated that the C391Y mutation in human VAChT is essential for vesamicol binding. The orthologous mutation in UNC-17 is C370Y and exists in the unc-17(md414) mutant, which also has weak reduction-of-function phenotypes. Without knowing that human VAChT (C391Y) fails to interact with vesamicol, the naïve expectation is that unc-17(md414) would also be hypersensitive to compound Ia. However, it was found that unc-17(md414) is partially resistant to the effects of both compound Ia and vesamicol (p=0.001). Together, these chemical-genetic interactions indicate that compound Ia likely inhibits UNC-17 function.

Example 6 Bioactivity Profile of Exemplary Compound Ia

Compounds of the Formula (I) fully inhibit egg-hatching of the mammalian parasites H. contortus and C. oncophora at mid-micromolar concentrations (30-60 uM) and inhibit dog heartworm movement with an ED₅₀ in the low micromolar concentration range (2-10 uM) in ex-vivo assays. The compounds have little effect on HEK293 cell proliferation at any concentration tested as shown above in FIG. 2 .

A dose-response analysis in fish of three compounds of Formula (I) showed that the molecules induce cardiac defects in the low micromolar range. However, exemplary compound Ia itself has the weakest cardiac defects of the three compounds, while compound Ie (which has relatively weak effects in worms), induced the strongest cardiac defects.

Example 7 Structural-Activity Relationship (SAR) of Compounds of Formula (I) and Bioactivity Profile

A preliminary first round of SAR analyses using analogs of exemplary compound Ia was performed and the analogs' ability to induce locomotory defects in wild type fourth-stage C. elegans larvae (L4s) was tested. As expected, most analogs induced weaker locomotory phenotypes compared to compound Ia, the exception being compound Ib (Table 1). Using this information, new analogs of exemplary compound Ia were synthesized and/or purchased, and the most potent analog to date, compound If (Table 1) was identified.

Table 1 shows the preliminary SAR study conducted on compounds of Formula (I), where activity indicates the percent of animals that show coiled and/or paralysis after 60 min on solid substrate with a “drug” concentration 60 μM. If phenotype is observed, a dose-response analysis is performed. Exemplary compound Ia has an EC₅₀ of 12.8 μM.

Example 8 Structural-Activity Relationship (SAR) of Compounds of Formula (I) and Bioactivity Profile

Table 2 shows the extension of the SAR study conducted on compounds of Formula (I), where data are the % of L3 animals demonstrating abnormal locomotory behaviour on solid agar (MYOB medium+E. coli OP50 food source) containing the indicated compound of the Application at 3.75 uM, 15 uM or 60 uM (as noted). 20 animals were scored after either 1, 6 or 24 hours of exposure to drug (as indicated) per replicate. Abnormal locomotory phenotype (phene) is defined as animals demonstrating 1 or more of the following phenotypes: jerky locomotion, flaccid paralysis, coiling. Data are reported as the mean % of animals with abnormal phenotype over 4 independent replicates.

While the present application has been described with reference to what are presently considered to be the preferred examples, it is to be understood that the application is not limited to the disclosed examples. To the contrary, the present application is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.

TABLE 1 Compound R² L—R¹ Activity (%) Ia NO₂

90% +/− 5.4 Ig F

11% +/− 5.6 Ih Cl

2% +/− 1.9 Ii H

19% +/− 8.1 Ik CN

61% +/− 5.0 Ib NO₂

87% +/− 3.7 Im NO₂

0.00 If NO₂

98% +/− 1.9 Ip CN

41% +/− 7.4 Iq NO₂

6% +/− 3.2 Ic NO₂

71% +/− 2.1 Id NO₂

0.00 Ie NO₂

19% +/− 8.0 Vesamicol — — 6% +/− 0.1 Piperazine — — 0.00

TABLE 2 % any phene 1 h (conc.) % any phene 6 h (conc.) % any phene 24 h (conc.) Compound I.D. 3.75 15 60 3.75 15 60 3.75 15 60 Control 0 0 0 0 0 0 0 0 0 Ia 42.5 91.0 100.0 22.5 64.2 100.0 0.0 34.2 80.0 Ic 29.2 95.9 100.0 8.3 80.8 100.0 0.0 89.2 100.0 Id 19.2 65.5 95.8 5.8 36.7 100.0 0.0 45.0 95.8 Ie 15.0 45.0 94.2 0.0 56.7 80.8 0.0 0.0 57.5 Ib 3.3 97.5 100.0 0.0 93.3 100.0 0.0 73.3 100.0 Im 0.0 7.5 44.2 0.0 3.3 30.8 0.0 0.0 0.0 Ig 21.7 44.2 95.0 0.8 9.2 79.2 0.0 4.2 66.7 Ih 0.0 27.5 55.7 0.0 0.0 5.8 0.0 0.0 10.8 Ii 16.2 39.2 84.2 0.0 0.8 44.2 0.0 0.0 20.5 Ir 43.3 97.5 100.0 9.2 88.3 100.0 0.0 30.0 100.0 Is 3.3 17.5 100.0 0.0 15.0 62.5 0.0 0.0 13.3 Ik 8.3 45.8 91.7 15.0 37.5 98.3 0.0 23.3 100.0 Ip 0.8 29.2 91.3 0.0 10.0 42.5 0.0 0.0 62.5 Iq 0.0 4.2 32.5 5.0 29.2 36.7 0.0 0.0 4.2 Iw 5.0 14.2 37.5 0.0 1.7 19.2 0.0 0.0 3.3 Ix 89.2 100.0 100.0 100.0 100.0 100.0 66.7 100.0 100.0 Iy 22.5 77.5 100.0 0.0 35.0 99.2 0.0 0.0 58.3 Iz 60.8 96.7 99.2 0.0 6.7 85.0 0.0 0.0 14.2 Iaa 45.0 89.2 100.0 6.7 27.5 97.5 0.0 0.0 69.2 Ibb 27.5 92.5 100.0 5.0 75.8 100.0 0.0 13.3 100.0 Icc 46.7 93.3 100.0 0.0 0.8 33.3 0.0 0.0 0.0 Idd 92.5 100.0 100.0 86.7 100.0 97.5 34.2 98.3 97.5 Iff 24.2 98.3 100.0 6.7 28.3 51.7 0.0 0.0 0.0 Igg 25.8 80.0 100.0 0.0 68.3 100.0 0.0 0.0 100.0 Ihh 4.2 70.0 100.0 0.0 1.7 53.3 0.0 0.0 5.0 Iii 81.7 96.7 100.0 14.2 94.2 100.0 0.0 77.5 100.0 vesamicol 0.0 0.0 11.7 0.0 0.8 1.7 0.0 0.0 0.0 

The invention claimed is:
 1. A method of treating or preventing a nematode infection comprising administering an effective amount of a compound of Formula (I) and/or a pharmaceutically acceptable salt and/or solvate thereof, to a subject in need thereof, wherein the compound of Formula (I) is selected from


2. The method of claim 1, wherein the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.
 3. The method of claim 1, wherein the compound of Formula (I) is

or a pharmaceutically acceptable salt and/or solvate thereof.
 4. The method of claim 1, wherein the infection is an infection of a nematode of a species selected from Cooperia oncophora, Haemonchus contortus, and Caenorhabditis elegans.
 5. The method claim 1, wherein the subject is selected from a mammal, a bird, a plant, a seed, and soil.
 6. The method of claim 1, wherein the subject is selected from a plant, a seed and soil and the compound is administered pre-planting, post-planting, as a feed additive, a drench, an external application, a pill and/or by injection.
 7. A method of treating or preventing a disease, disorder or condition arising from a nematode infection comprising administering an effective amount a compound of Formula (I) and/or a pharmaceutically acceptable salt and/or solvate thereof to a subject in need thereof wherein the compound of Formula (I) is selected from


8. The method of claim 7, wherein the compound of Formula (I) is selected from

or a pharmaceutically acceptable salt and/or solvate thereof.
 9. The method of claim 7, wherein the compound of Formula (I) is

or a pharmaceutically acceptable salt and/or solvate thereof.
 10. The method of claim 7, wherein the infection is an infection of a nematode of a species selected from Cooperia oncophora, Haemonchus contortus, and Caenorhabditis elegans.
 11. The method claim 7, wherein the subject is selected from a mammal, a bird, a plant, a seed, and soil.
 12. The method of claim 7, wherein the subject is selected from a plant, a seed and soil and the compound is administered pre-planting, post-planting, as a feed additive, a drench, an external application, a pill and/or by injection.
 13. A method of treating or preventing a nematode infection comprising administering an effective amount of a composition comprising a compound of Formula I as defined in claim 1, or a pharmaceutically acceptable salt and/or solvate thereof, to a subject in need thereof.
 14. The method of claim 13, wherein the subject is selected from a plant, a seed and soil and the composition is administered pre-planting, post-planting, as a feed additive, a drench, an external application, a pill and/or by injection.
 15. The method of claim 1, wherein the subject is a human.
 16. The method of claim 7, wherein the subject is a human.
 17. A method of treating or preventing a disease, disorder or condition arising from a nematode infection comprising administering an effective amount a composition comprising compound of Formula (I) as defined in claim 7, or a pharmaceutically acceptable salt and/or solvate thereof, to a subject in need thereof.
 18. The method of claim 17, wherein the subject is selected from a plant, a seed and soil and the composition is administered pre-planting, post-planting, as a feed additive, a drench, an external application, a pill and/or by injection. 